DE2609533C3 - Process for the extraction of active substances, in particular of heteroside esters of caffeic acid, as well as medicaments containing these compounds - Google Patents

Description

whereby /;; is an integer from 0 to 5 contains.

2. Process for extracting a heteroside ester of the formula

HO OH

HO - <ζ S - X - O - (glucose rhamnose) - OOC - CH = CH - <ζ y - OH where X is a group - (CH ₂ ), -. where η is an integer from 1 to 6. or

OH where m is an integer from O to S,

characterized in that plants of the families Scrophulariaceae, Orobanchaceae, Labiatae or \ canthaceae soaked in an alcohol such as ethanol or methanol, the alcoholic 4 » Solution wins, the alcohol evaporates, the residue obtained is taken up again in water and extracted with ethyl acetate, and that the fraction soluble in ethyl acetate is recovered

The invention relates to new drugs and the production of their active ingredients by extraction of -n Plant materials.

It is already known that heteroside esters of caffeic acid can be extracted from certain plants can. So one has z. B. already produced the heteroside ester, which differs from caffeic acid, rhamnose, v> of glucose and derived from dihydroxyphenylethanol. This has been referred to as a verb-fascoside, for one To give an indication of the particular plant from which it was extracted, namely Verbascum sinuatum. The investigations carried out with regard to these products have, however, been limited to structural investigations and none have been carried out so far Property of these compounds found that make them suitable for any technical application would. M)

The present invention is based on the discovery that heteroside esters of caffeic acid, obtained from various plants and in particular from plants of the Scrophulariaceae families, the Orobanchaceae. the Labiatae or the Acanthaceae tr. extract can show interesting phartnacological properties that they are responsible for the formation of Make medicinal products suitable.

According to the invention, the active ingredients, in particular heteroside esters of caffeic acid, can be used and which are suitable for pharmaceutical use by the following process in which one plants, namely Tumifloralae plants, preferably Scrophulariaceae, Orobanchaceae, Labiatae, Acanthaceae, or analogous plants, soaked in an alcohol that wins the alcoholic solution Alcohol evaporates, the remaining aqueous phase is subjected to extraction with ethyl acetate and the the fraction which is soluble in ethyl acetate is recovered. In particular, ethanol or methanol can be used as the alcohol be used.

Among other things, it is preferable to subject the plants to be used to prior degreasing, preferably with petroleum ether.

This treatment is advantageously carried out with degreasing the residue of the alcoholic solution completed, which is done before extraction with ethyl acetate. For this purpose a degreasing Solvent, /. B. ethyl ether or petroleum ether is used. So you can z. B. in particular as follows Procedure: The entire plant is dried, ground and then mixed with petroleum ether in one Degreased Soxhlet device. The received »; Residue

or grains which has been freed of petroleum ether, is soaked in ethyl alcohol of 80 ⁰ C or so leached The alcoholic solution is concentrated in vacuo until the alcohol is removed, the remaining aqueous liquid is then decolorized by treatment with ether and this is removed It is then treated with ethyl acetate. The ethyl acetate phase is dehydrated and concentrated to dryness. In this way, a creamy white powder is obtained as residue, which is the extract prepared according to the invention

According to a preferred embodiment of the invention, in particular plants of the family Orobanchaceae used are the parasitic phanerogams that are stripped of chloroform. Under these plants are in particular the species of the genus Orobanche and those of Phelypaea preferred These species have the advantage that they are relatively rich in the derivatives of caffeic acid and that they therefore give good extraction yields.

Nevertheless, the plants are also of the Scrophulariaceae family, such as Verbascum, and those of the Acanthaceae family such as B. Acanthus, of interest because they are often more readily available are. In this case, in particular, the method is advantageously carried out by purifying the extract in the aqueous medium, in particular on Ceüufose, and then by a renewed extraction by with

ι ο methanol-added ethyl acetate completed

The process according to the invention makes it possible in particular to obtain extracts which are hereinafter referred to as "Polyphenol extracts" are called. The analysis has shown that these are Mixtures are, which contain essentially different isomers, which heteroside esters of caffeic acid derivatives represent a heteroside of a dihydroxyphenylalkanol, and in particular the following Have formula:

OH

- O - (glucose rhamnose) - OOC - CH = CH - <f

OH

Here X means the group:

- (CH ₁ ), -. where η is an integer from 1 to 6

- CH — iC H ₂ ) ", where m -; me is an integer from 0 to 5.
OH

Certain of these isomers can advantageously be separated off from this mixed extract by completing the process by one or more recrystallizations in water.

The relative proportions of the isomers in the Polyphenol extract may vary depending on the treatment conditions and on the exact nature of the plants used vary.

The invention also relates to compounds obtained by this process.

Among the compounds of formula I, preferred compounds according to the invention are those Heteroside esters of caffeic acid derivatives of heterosides of dihydroxyphenylethanol and of dihydroxyphenylglycol of the formulas:

HO
HO— <f V-CH-CH, - () - (Glucose-Rhamnose) - OOC - CH = CH- <f> —OH (II)

S = / j

OH
and

HO OH

HO— <fV — CHj — CH ₂ -O- (Glucose ^ Rhiimnose) - OOC - CH = CH - <^ V-OH (III)

Particularly important amounts of these compounds are found in the polyphenol extracts of Acanthaceae or Sorophulariaceaen for a compound of the formula III, which is given the name Verbascosid has given, and in the polyphenol extracts of the Orobanchaceaen for a compound of the formula H, the the name Orobanchosid was given. You can by crystallization from the aqueous solutions of the Polyphenol extract can be isolated and purified by recrystallization from water or alcohol.

Other isomers of the caffeic acid esters according to the invention can be obtained by using the Verbascoside or orobanchoside in weakly acidic medium or their mixtures or directly

Polyphenol extract heated. Heating in the weakly acidic medium of the orobanchoside, preferably by Cooking at a pH of 6 to 6.8 results in an isomer of the general formula given above, which has been called "Isoorobanchosid". In the same way, heating the verbascoside leads to the weakly acidic medium under analogous conditions to an isomer of the same general formula that has been called "isobescoside".

All isomers isolated in this way show, like the polyphenol extract, pharmacological properties, which turn them into active ingredients that are suitable for the formation of drugs. You still have the advantage that they can easily be obtained either separately or in a mixture by the process according to the invention crystalline state can be obtained.

The invention therefore also takes into account medicaments which contain at least one heterositol ester as an active ingredient contain caffeic acid. In particular, those drugs are considered that at least a heteroside ester of a caffeic acid derivative of a heteroside of a dihydroxyphenylalkanol, preferably of the above formula I or in particular the above formula II, or the at least one Contain active ingredient, as made by the process according to the invention with different operating variants Plants of the family of the Orobanchaceaen, the Scrophu-Iariaceaen, the Labiataen or the Acanthaceaen can be extracted.

These active ingredients show in particular a potentiating Effect for dihydroxyphenylalanine or dopa. They can be used in combination with dopa for advantage whose various therapeutic indications are administered. The main current indication is the treatment of Parkinson's disease

It is known that dopa, especially in the levorotatory form or as L-dopa, is currently known as considered the most effective drug against Parkinson's disease. Unfortunately, it has However, this drug has a number of side effects that affect cardiovascular function and the intensity of which can lead to intolerance. It is also known that this Side effects are due to the presence of a large amount of dopamine, which is from a Extracerebral activity of dopa decarboxylase originates, which causes the decarboxylation of dopa. For the treatment of Parkinson's disease, it is therefore of great interest to have one with the L-Dopa To be able to connect inhibitor for the extracerebral dopadecarboxylase. The active ingredients according to the invention have now been shown to be inhibitors of this type. Furthermore, di ·; active ingredients according to the invention the advantage that they have longer lasting and more effective activity than the original synthetic chemical Connections such as B. the caffeic acid itself, and that they have no side effects.

The active ingredients according to the invention also have other pharmacological properties. In particular Hje polyphenol extracts have a ^ -blocking Activity that also increases her interest in the treatment of Parkinson's disease. One avoids hence the parallel administration of ^ -blocking drugs with dopa-potentiating drugs, as is customary in the treatment of Parkinson's disease, or at least that too the doses to be administered. The activities required can be found in a single product of vegetable origin combined and the activity lasts longer and the agent is better tolerated than that Products that are used in the classic way.

The invention therefore also relates to pharmaceutical preparations, in particular for the treatment of Parkinson's disease, which has a heteroside ester of caffeic acid or, in particular, one after the Process according to the invention obtained active ingredient together with dihydroxyphenylalanine, which is preferably is present in left-handed form.

The invention is illustrated in the examples, which illustrate the preparation of the active ingredients and their constitution and their pharmacological properties are described.

example 1
Production of a polyphenul extract

From plants of the genus C-: jbanche, in particular of the species Orobanche hederae Dnby and Orobanche rapum Thuill, a polyphenol extract is produced.

The whole plants that have been collected in June / July are sent immediately after collecting Dried in a ventilated place and urn away from light

400 g of the whole dried plants are after

grinding to a semi-fine powder with 61

jo petroleum ether (40 to 60 ⁰ C) for 10 to 12 hours degreased and depigmented or decolorized in a Soxhlet device.

The plants thus obtained powder is dried and then soaked in the air at a temperature of 4O ⁰ C J5 with 5x21 ethanol at 80 ⁰ C under mechanical stirring or leached.

After filtering off the spent grains or the extracted material, the 101 become the alcoholic Solution mixed with 20 ml of a 10% aqueous sodium metabisulphite solution, whereby the oxidation the phenolic active ingredients is reduced.

These solutions are kept in the refrigerator overnight

leave (4 ⁰ C) and then filtered and concentrated by the alcohol is stripped off in a rotary evaporator under vacuum at moderate temperature (30 "C)..

The extract, which has now become aqueous, is in turn defatted and decolorized by using a Mixture of the following is stirred:

Ethyl ether (freed from its peroxides)
Petroleum ether (40 to 6O ⁰ C).

The respective volumes are 2.5 / 0.5 1.

This aqueous extract, which has been freed from traces of ether, is then by adding more Water made more watery and with 181 freshly distilled ethyl acetate (which takes the active ingredients with it) extracted into decanting ampoules, which are moved mechanically.

The ethyl acetate phases are dried over sodium sulfate (pure, dry, from Prolabo) and evaporated to dryness in vacuo until the organic solvent has completely disappeared.

In this way, 40 g of a product in the form of μ becomes a light powder with a creamy white color obtained, which is the d ^ n polyphenol extract.

Yield: 10% by weight, based on the dry plant.

Example 2
Production of the orobanchoside

The polyphenol extract prepared according to Example I is distilled warm in such an amount Dissolved in water so that a 20% solution of the extract is obtained. After the solution has cooled down At ambient temperature, pink noodles are formed Crystals with a light beige color. These are collected and they represent the raw orobanchoside.

The orobanchosid crystal are made by dissolving in absolute ethanol in the warmth and in the presence of some powdered biochar of the type »Norit« (Prolabo) cleaned. The solution is provided by a small, with cellulose (300 M. N.) filled column with a height of 1 cm and a diameter of 2 cm, the previously washed with water and with ethanol, filtered.

The alcoholic riiiiiii witi) in a vacuum in one Rotary evaporator concentrated until / around the occurrence of an exercise. Put in a refrigerator (at 4 C) left to crystallize. The crystals obtained are centrifuged off, dried and once or twice in recrystallized from absolute ethanol.

Yield of purified orobanchoside: 2% by weight. based on the dry plants.

The product obtained has the following physicochemical Characteristics:

a) Physical properties

Melting point: determined on a Kofier · Hank

Softening begins at 196 to 200 ° C

Clear melting: 209 ± ■ C;

Solubility:

insoluble in cold water

sufficiently soluble in wallowing water

soluble in alcoholic and alcoholic-aqueous solutions

insoluble in ether. Chloroform and benzene.

b) elemental analysis

Found: C = 54.50%. H = 5.78%.
C = 54.62%. H = 5.87%,
Calculated: C = 54.37%. H - 5.62% for Ch. H, "O".

c) Analysis by thin layer chromatography:

Adsorbent:
Solvent:

Duration of the hike:
Development funds:

Layer of Merck cellulose on an aluminum foil
2% aqueous acetic acid solution

1 h 30 min

UV light and diazotized p-nitroaniline (yellow-beige color) 0.43 to 0.44.

d) Spectral analysis:

UV spectrum (in methanol):

Maximum 223- (233) - (246) -291 -335 Minimum 265

Infrared spectrum (in KBr tablet) - location of the bands in cm ~ ':

3420-2960-2935-1720-1605-1520-1450-1380-1300-1270-1130-1060-1040-1020-980-850-810-780-570. Nuclear Magnetic Resonance Spectrum (NMR):

6 aromatic protons 6.4 ppm <<5 <7.2 ppm 2 protons of a trans double bond J = 15 Hz (δ = 6.2 ppm <5 = 7.5 ppm)

3 methylprolones J = 6Hz Λ = 1.05 ppm
I proton ό = 5.03 ppm (CiRhamnosyl)
I proton ό = 4.58 ppm (C ₁ Glucosylj (J = 7 Hz)
+ 13 or 14 protons between 3 ppm and 5.1 ppm.

e) Chemical formula

On the basis of the analytical results given above and the investigation of the products separated by thin-layer chromatography on cellulose after alkaline hydrolysis of orobanchoside, it can be concluded that this product is formed from the following:

1 molecule of caffeic acid
I molecule of glucose
I rhamnose molecule

1 molecule ß- \ lydroxv / ifdihydroxy3,4-phenylethyl alcohol !. according to formula II.

Fi c ι s ρ ie I i

Production of the isoorobanchoside

The orobanchoside prepared according to Example 2 is in an aqueous medium at a slightly below 7, namely in the region of 6.5. lying pH and the solution is kept boiling for i Ii.

The solution is then allowed to cool. It Crystals separate and are centrifuged off and purified as in Example 2. The received Product, d. H. the isoor-jbanehosid. isi an isomer of Orobanchosids of Example. '. which has the same general formula II.

Example 4
Production of the verbascoside

The verbascoside is extracted from mother liquors,

into those after the separation of the crystallized orobanchoside according to Example 2, starting from a Polvnhenol-F.xtraki provided according to Example 1. lag behind and it is dissolved in water.

The mother liquors are in a refrigerator

4, (4 ° C) left to stand in a beaker with covered with a watch glass. which allows some evaporation. After about a week Crystals appear in the beaker, which are clumped together in the form of small spheres.

> (i These crystals are centrifuged off and, after redissolving them in water at ambient temperature (solution with 25 to 30%), allowed to crystallize in the refrigerator. After 48 h the verbascoside crystallizes

The process is repeated until a single spot of the verbascoside is obtained on the chromatograms.

The product obtained has the following physicochemical properties:

a) Physical properties

Melting point (Kofier bench)

Soften at approximately 158 to 160 ° C clear melting at about 170-180 ° C Solubility:

soluble in cold water

soluble in alcohol

insoluble in ether, chloroform and benzene.

b) chromatography

The product is analyzed by thin layer chromatography under the same conditions as the orobanchoside analyzed in example 2.

The stain corresponding to the verbascoside is discolored by diazotized p-nitroaniline to yellowish-brown, which is fringed with a mauve color.
Ri = · 0.57 (solvent: 2% acetic acid).

c) Spectral analysis

UV spectrum (in methanol):

A Maximum: 223 - (234) - (246) -291-Ji3

Minimum: 265

Infrared spectrum (in KBr tablets) location of the bands in Lin ':

3400-2920-1640-1595-1510-1440-

1 170-1260-116O- I 11 5- 1060-1030 -

980-810-780
Nuclear resonance / .spcklrum (NMK):

6 aromatic protons (6.4 <Λ <7.2 ppm)

2 protons of a fans double bond
I = 15 H /

(Λ = 6.25 ppm and ό - 7.55 ppm)

3 methyl protons J = 6 H / ti = 1.05 ppm
1 proton Λ = 5.12 (Ci Rhamnosyl)

I proton Λ = 4.40 ppm (C, glucosyl) J = 7 M /.
+ 14 protons between 2.6 ppm and 5.0 ppm.

The results of the analysis of the hydrolysis products of the verbascoside / eigen that this differs from the similar components, namely glucose. Rhamnose and caffeic acid, such as the orbanchoside and of /J(Dihydroxy-3.4-phenyl) ethanol and that it derives the Formula III has.

Example 5
Manufacture of isobescoside

The verbascoside produced according to example 4 is heated according to example 3 in an aqueous, slightly acidic medium, whereby the isobascoside is obtained \ Utrvl AtK ΑΊ & itlnw>Kt> illiiDmoii.n ΓητηιοΙ Ml h ^ i

Example 1 6

A polyphenol extract from Orobanche is made as in the Example 1 prepared and prepared by thin layer chromatography on cellulose with 2% aqueous acetic acid analyzed. After the chromatogram has been developed using UV light and diazotized p-nitroaniline, the following can be distinguished:

in strong concentration

Verbascoside: Rf = 0.57

Orobanchoside: Rf = 0.43
in very weak concentration:

Isobescoside: Rf = 0.24-0.25

Isoorobanchoside: Rf = 0.18-0.20
in trace amounts:

Caffeic acid: Rf = O ₁ IO.

The physicochemical properties of the polyphenol extract are as follows:

Solubilities:

Instantly soluble in ambient temperature water to a medium concentration solution (after 1 h some orobanchoside crystals separate in the cold)

soluble in alcohols and ethyl acetate
insoluble in ether, chloroform and benzene.
UV spectrum (in methanol):

the same as the caffeic acid ester
A maximum: 221- (248) -29l-335
Minimum: 266

Example 7

Plants of the genus Verbascum are dried and ground. Then they are according to Example 1 treated by degreasing, extraction with alcohol, degreasing and extraction with ethyl acetate. It will be like in the Proceed in Example I until the ether-acetate extract that remains after the ethyl acetate has evaporated is obtained.

This extract is then dissolved in water and passed through a column of cellulose and purified by elution with a 10% aqueous acetic acid solution. The aqueous phase is then extracted through ethyl acetate to which 3% methanol has been added.

After drying on anhydrous sodium sulphate and evaporation to dryness, it becomes a powdery one cream-colored residue obtained. The analysis shows that it is largely the verbascoside of Formula III acts.

Example 8

Toxicological investigation

The active ingredients of plant origin produced according to the invention have no toxicity and they do are very well tolerated.

For the polyphenol extract obtained according to Example I. the results of the toxicological examination are as follows:

acute toxicity:

DL. 50> 6 g / kg per os in the mouse
Dl. 50> 6 g / kg ip in the mouse

The "traction" test is positive 2 hours after ingestion of the product in a high dose (1 g / kg ip). This Tijt consists essentially in the fact that one can see the effect of the p .. ", i ,, i ,; _r o..r, i " _n ΚΛ. ..- i. "i." ...... "- .. u -ι" - γ "" ι ...: ι. _vc julou and Courvoisier: Psychotropic drugs, Milan (1957), 373.

Chronic toxicity: at 500 mg / kg body weight of the rat for eight weeks there are none Ads noted.

Analogous results are obtained with each active ingredient prepared according to Examples 2 to 7.

Example 9
Potentiating activity for L-dopa

The studies on the potentiation of the antiparkinsonian activity of L-Dopa are in the In a way, that one carried out the antagonism of these two substances against that in the mouse Investigated tremors induced by oxotremonine.

The trembling movements are recorded graphically.

In any case, the following protocol is followed:

Time 0

= the active ingredient is administered orally
Time 20 min

= L-Dopa is administered orally
Time 40 min

= 500 y / kg oxotremonin are i. p. administered

The tremors become 45 minutes
Administration of the oxotremonine registered.

Examination of the results shows the same antioxotremonin activity in:

L-dopa alone = 500 mg / kg per os,

or any of the following combinations of active ingredient + L-dopa:

1) Orobanche polyphenol extract (Example 1) = 50 mg / kg per os

(.-Dopa = 50 mg / kg per os

2) Orobanche polyphenol extract (Example I) = 100 mg / kg orally

I.-Dopa = 25 mg / kg per os

3) orobanchoside (example 2) = 100 mg / kg per os
L-Dopa = 100 mg / kg per os

4) Vcrbaskosid (Example 4) = 100 mg / kg per os
L-Dopa = 100 mg / kg orally.

The mechanism of action for the potentiation of L-dopa is shown in vitro under the following conditions examined:

Substrate

= I.Dopa
enzyme

= powdered guinea pig kidney grated in acetone, produced at very deep temperature

enzymatic control inhibitor
= Caffeic acid.

With the polyphenol extract obtained according to Example 1, the inhibiting effect on dopadecarboxylase is negative under the test conditions (pH = 6.8). On the other hand, when the pH value in the intestine is alkaline at ¹ 37 ° C, this extract releases caffeic acid, which is active. This could explain the inhibitory activity against dopa decarboxylase, assuming that it is formed in the organism after hydrolysis of the extract.

Example 10

Cardiovascular effect

Central pressure

Carotid artery pressure is measured in male rats from the Wistar family.

The administration of 50 mg / kg of polyphenol extract of Example 1 by the intravenous route is effective without Change in heart rate a free and temporary high blood pressure. - isolated heart chamber the rat.

The same polyphenol extract from Orobanche lifts at a dose of 50 mg in a volume of 40 ml all positive chronotropic and inotropic effects caused by 0.5 g of isoprenaline when testing the isolated Ventricle of the rat. This test consists of getting the rhythm and measures the contractions of an isolated heart chamber or an isolated auricle which survived in a nutrient liquid is held. This technique is described in J. Physiol. (1926) - 61,547.

The extract therefore has a yes-blocking effect Effect.

Antihypertensive activity

An experimental one realized by rhenal ischemia Hypertension (Groilman technique: Proc. Soc.

Biol. Med. 1944, 57, 102 to 104) is increased by two points by daily administration of 300 mg / kg per os of the polyphenol extract of Orobanche, prepared according to Example 1, diminished.

The drop in arterial pressure, which begins three days after the start of treatment, persists during sustained for the entire duration of the treatment. 10 days after the end of treatment, the arterial goes Pressure back up to its initial value.

It should be noted that the extract does not modify the arterial pressure of normally toned animals.

Anti-flicker activity

The same polyphenol extract does not antagonize that in the Lawson method mouse generated chloroform fibrillation.

Example 11
Aiialgeiiscne Akiiviiäi

The specified i. p. doses of the polyphenol extract obtained according to Example I of Orobanche raising the pain threshold in two techniques that involve thermal stimulation (Test of the hot plate) and chemical stimulation (test with phenylbenzoquinone) can be used.

Heating plate:

according to the method of Woolfe (G.) - MacDonald (A. D.) in J. Pharmacol, exptl. ther. (1944), 80,300.

This test is based on the following principle: A mouse placed on a plate heated to 56 "C reacts 5 to 6 seconds by licking the front paws or jumping. The administration of a Analgesic has the effect of increasing the time it takes for this response to occur.

The results obtained are summarized in the following table, which shows the percentage enlargement the time it takes for it to appear in response to the pain.

500 mg / kg
1 g / kg ip i.p. M) min 60 min W) min 92
170 80
130 50
100 Dose:
Dose:

Phenylbenzoquinone:

according to the method of Sigmund: J. Pharmacol, exptl. ther. (1957), Π 9.184.

This test is based on the following principle: After the prophylactic administration of an analgesic an alcoholic solution of phenylbenzoquinone is injected, which causes the animal to twist. the Analgesics mitigate or inhibit this twisting syndrome.

The results obtained are summarized below as a percentage reduction:

Line in min

5 10 15 20 25 30

Dose: 500 mg / kg ip
Dose: 1 g / kg ip

100 78 69 74 67 65 i00 94 91 85 S3 85

Used in the experimental conditions and in two At doses, the examined extract showed strong analytical activity.

Example 12
Anti-inflammatory activity

According to the method of Winter - Proc. Soc. Exp. Biol. med. (1962), 111,544-547.

Injection of a kaolin suspension into the sole tendon skin of the rat's hind paw results in a Inflammatory reaction, which can be reduced by anti-inflammatory substances.

In the following table those with derr> Polypheiiol extract of Example 1 obtained results compiled. They are given as a percentage reduction in swelling:

I 2 .1 4 5 (i

Dose: 500 mg / kg i.p.

IO 42 25 35 20 10

Under the test conditions, the tested product showed anti-inflammatory activity.

Example 13
Psychotropic effects

The product investigated is the polyphenol extract from Orobanche, obtained according to Example I.

Comprehension and motility

Swiss mice are exposed to the Boissier method (Therapy [1962], 17, 1225 to 1232) Mobility and comprehension examined.

The results obtained are expressed as the percentage variation in perception and motility, based on the control animals, compiled in the following table:

Cans 1 g / kg 50 (1 mg / kg 1. P- I.p.

Comprehension
Motility

- 34%

- 33%

- 79%

- 65%

At the doses examined, the product reduced comprehension and motility.

Examination of the reflexes

Traction test:

Test positive (no effect of the product) with a Dose of 1 g / kg i.p. p.

Rotation test according to the Gross technique (Schw. Med.wshr. [1955], 85, 305).

This test determines the period of time over which a mouse is in equilibrium on a horizontal shaft that rotates around itself can hold: test positive (no effect of the product) at a dose of 1 g / kg i.p. p.

Schi afpo ten door

The potentiation of the anesthesia is a test that is used to test a general sedative effect. ^p Sl'bstäMZ, which does not produce the Schlilf itself, can, in combination, extend the duration of hypnotic activity.

The anesthesia is increased in male Swiss mice by the technique of euthanasia determined (technique by Courvoisier in Arch. int. pharmacod. ther. [1952], 92, 305: Sleep, through a subhypnotic dose has been induced. is by the effect of a pre-treatment by a Psycholeptic prolonged).

500 mg / kg of the compound administered intraperitoneally. bring compared to that Sleep time of the control animals resulted in an increase in sleep time of 43%.

The tested active ingredient diminished without exposure on the reflexes the comprehension and the motility and it potentiates a barbiturate narcosis. What proves a psycholeptic activity.

Example 14

Capsules suitable for human therapy. have a dose of 50mg of polyphenol extract Orobanche according to Example 1 per capsule.

This extract can be administered in daily doses from 1 to ■> mgKg in connection with daily doses of L-D01.L from 6 to 30 mg / kg.

For example, one can tell a patient who has Parkinson's disease. 200 mg extract per Administer in 3 or 4 doses daily in conjunction with a dose of 1 g L-Dopa per day.

Claims (1)

ι 2 claims: 1. Medicament, characterized in that there is at least one heteroside ester of the formula HO OH UO - <f \ --X - O - (glucose rhamnose) - OOC - CH = CH - <f ^ --OH wherein X is a group - (CHi) _n -, where η is an integer from 1 to 6, or - CH- (CH ₂ ) ,, - I.
OH